Metabolic engineering to simultaneously activate anthocyanin and proanthocyanidin biosynthetic pathways in Nicotiana spp

[EN] Proanthocyanidins (PAs), or condensed tannins, are powerful antioxidants that remove harmful free oxygen radicals from cells. To engineer the anthocyanin and proanthocyanidin biosynthetic pathways to de novo produce PAs in two Nicotiana species, we incorporated four transgenes to the plant chassis. We opted to perform a simultaneous transformation of the genes linked in a multigenic construct rather than classical breeding or retransformation approaches. We generated a GoldenBraid 2.0 multigenic construct containing two Antirrhinum majus transcription factors (AmRosea1 and AmDelila) to upregulate the anthocyanin pathway in combination with two Medicago truncatula genes (MtLAR and MtANR) to produce the enzymes that will derivate the biosynthetic pathway to PAs production. Transient and stable transformation of Nicotiana benthamiana and Nicotiana tabacum with the multigenic construct were respectively performed. Transient expression experiments in N. benthamiana showed the activation of the anthocyanin pathway producing a purple color in the agroinfiltrated leaves and also the effective production of 208.5 nmol (-) catechin/g FW and 228.5 nmol (-) epicatechin/g FW measured by the p-dimethylaminocinnamaldehyde (DMACA) method. The integration capacity of the four transgenes, their respective expression levels and their heritability in the second generation were analyzed in stably transformed N. tabacum plants. DMACA and phoroglucinolysis/HPLC-MS analyses corroborated the activation of both pathways and the effective production of PAs in T0 and T1 transgenic tobacco plants up to a maximum of 3.48 mg/g DW. The possible biotechnological applications of the GB2.0 multigenic approach in forage legumes to produce "bloatsafe" plants and to improve the efficiency of conversion of plant protein into animal protein (ruminal protein bypass) are discussed.This work was supported by grants BIO2012-39849-C02-01 and BIO2016-75485-R from the Spanish Ministry of Economy and Competitiveness (MINECO) (http://www.idi.mineco.gob.es/portal/site/MICINN) to LAC and a fellowship of the JAE-CSIC program to SF. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Fresquet-Corrales, S.; Roque Mesa, EM.; Sarrión-Perdigones, A.; Rochina, M.; López-Gresa, MP.; Díaz-Mula, HM.; Belles Albert, JM.... (2017). Metabolic engineering to simultaneously activate anthocyanin and proanthocyanidin biosynthetic pathways in Nicotiana spp. PLoS ONE. 12(9). https://doi.org/10.1371/journal.pone.0184839Se018483912

Nurses' perceptions of aids and obstacles to the provision of optimal end of life care in ICU

Contains fulltext : 172380.pdf (publisher's version ) (Open Access

All-fiber processing of terahertz-bandwidth signals based on cascaded tapered fibers

Tapered single-mode fibers are employed to perform dynamic pulse shaping in a bandwidth of several terahertz. The transfer function of cascaded biconical tapers is controlled by introducing a phase shift into one of them through mechanical stretching. It is a simple and low-cost technique with potential to process signals with bandwidths as large as those allocated by standard optical fiber while introducing little degradation. Femtosecond pulses are shaped to prove the concept. (C) 2013 Optical Society of AmericaThis work was supported by the Generalitat Valenciana programs PROMETEO-2010-087 R&D Excellency and GV/2012/121 Grupos Emergentes.Mas Gómez, SM.; Palací López, J.; Pérez Millán, P.; Lechago Buendía, S.; Monzón-Hernández, D.; Martí Sendra, J. (2013). All-fiber processing of terahertz-bandwidth signals based on cascaded tapered fibers. Optics Letters. 38(23):4954-4957. https://doi.org/10.1364/OL.38.004954S495449573823Lacroix, S., Gonthier, F., Black, R. J., & Bures, J. (1988). Tapered-fiber interferometric wavelength response: the achromatic fringe. Optics Letters, 13(5), 395. doi:10.1364/ol.13.000395Villatoro, J., Monzón-Hernández, D., & Mejía, E. (2003). Fabrication and modeling of uniform-waist single-mode tapered optical fiber sensors. Applied Optics, 42(13), 2278. doi:10.1364/ao.42.002278Monzón-Hernández, D., & Villatoro, J. (2006). High-resolution refractive index sensing by means of a multiple-peak surface plasmon resonance optical fiber sensor. Sensors and Actuators B: Chemical, 115(1), 227-231. doi:10.1016/j.snb.2005.09.006Villatoro, J., Monzón-Hernández, D., & Luna-Moreno, D. (2004). In-line optical fiber sensors based on cladded multimode tapered fibers. Applied Optics, 43(32), 5933. doi:10.1364/ao.43.005933Wang, Z., Sone, H., Tsuji, Y., & Imai, M. (2007). Spectral Phase and Intensity Evolutions of Supercontinuum Generation in a Biconical Tapered Fiber. Optical Review, 14(2), 81-85. doi:10.1007/s10043-007-0081-6Lu, F., & Knox, W. H. (2004). Generation of a broadband continuum with high spectral coherence in tapered single-mode optical fibers. Optics Express, 12(2), 347. doi:10.1364/opex.12.000347Jedrzejewski, K. P., & Kosinski, A. (1997). Wide-band tapered directional coupler. Technology and Applications of Light Guides. doi:10.1117/12.285622Kawasaki, B. S., Hill, K. O., & Lamont, R. G. (1981). Biconical-taper single-mode fiber coupler. Optics Letters, 6(7), 327. doi:10.1364/ol.6.000327Rusu, M., Herda, R., Kivistö, S., & Okhotnikov, O. G. (2006). Fiber taper for dispersion management in a mode-locked ytterbium fiber laser. Optics Letters, 31(15), 2257. doi:10.1364/ol.31.002257Kieu, K., & Mansuripur, M. (2007). Femtosecond laser pulse generation with a fiber taper embedded in carbon nanotube/polymer composite. Optics Letters, 32(15), 2242. doi:10.1364/ol.32.002242Weiner, A. M. (2011). Ultrafast optical pulse shaping: A tutorial review. Optics Communications, 284(15), 3669-3692. doi:10.1016/j.optcom.2011.03.084Liu, X., Premkumar, A., & Madhukumar, A. (2008). Pulse shaping functions for UWK systems. IEEE Transactions on Wireless Communications, 7(5), 1512-1516. doi:10.1109/twc.2008.061060Weling, A. S., & Heinz, T. F. (1999). Enhancement in the spectral irradiance of photoconducting terahertz emitters by chirped-pulse mixing. Journal of the Optical Society of America B, 16(9), 1455. doi:10.1364/josab.16.001455Weiner, A. M., Heritage, J. P., & Kirschner, E. M. (1988). High-resolution femtosecond pulse shaping. Journal of the Optical Society of America B, 5(8), 1563. doi:10.1364/josab.5.001563Saperstein, R. E., Alić, N., Panasenko, D., Rokitski, R., & Fainman, Y. (2005). Time-domain waveform processing by chromatic dispersion for temporal shaping of optical pulses. Journal of the Optical Society of America B, 22(11), 2427. doi:10.1364/josab.22.002427Birks, T. A., & Li, Y. W. (1992). The shape of fiber tapers. Journal of Lightwave Technology, 10(4), 432-438. doi:10.1109/50.134196Kieu, K. Q., & Mansuripur, M. (2006). Biconical Fiber Taper Sensors. IEEE Photonics Technology Letters, 18(21), 2239-2241. doi:10.1109/lpt.2006.884742Cohoon, G. (2010). Enhancing sensitivity of biconical tapered fiber sensors with multiple passes through the taper. Optical Engineering, 49(3), 034401. doi:10.1117/1.3339906Ji, W. B., Liu, H. H., Tjin, S. C., Chow, K. K., & Lim, A. (2012). Ultrahigh Sensitivity Refractive Index Sensor Based on Optical Microfiber. IEEE Photonics Technology Letters, 24(20), 1872-1874. doi:10.1109/lpt.2012.221773

Discovery of the first dual GSK3β inhibitor/Nrf2 inducer. A new multitarget therapeutic strategy for Alzheimer's disease

The formation of neurofibrillary tangles (NFTs), oxidative stress and neuroinflammation have emerged as key targets for the treatment of Alzheimer's disease (AD), the most prevalent neurodegenerative disorder. These pathological hallmarks are closely related to the over-activity of the enzyme GSK3β and the downregulation of the defense pathway Nrf2-EpRE observed in AD patients. Herein, we report the synthesis and pharmacological evaluation of a new family of multitarget 2,4-dihydropyrano[2,3-c]pyrazoles as dual GSK3β inhibitors and Nrf2 inducers. These compounds are able to inhibit GSK3β and induce the Nrf2 phase II antioxidant and anti-inflammatory pathway at micromolar concentrations, showing interesting structure-activity relationships. The association of both activities has resulted in a remarkable anti-inflammatory ability with an interesting neuroprotective profile on in vitro models of neuronal death induced by oxidative stress and energy depletion and AD. Furthermore, none of the compounds exhibited in vitro neurotoxicity or hepatotoxicity and hence they had improved safety profiles compared to the known electrophilic Nrf2 inducers. In conclusion, the combination of both activities in this family of multitarget compounds confers them a notable interest for the development of lead compounds for the treatment of AD.R.L. thanks Instituto de Salud Carlos III co-financed by the European Regional Development's funds (FEDER) for a research contract under Miguel Servet Program (CP11/00165) and financial support from the European Commission-REA, People (Marie Curie Actions) FP7 under REA grant agreement n° PCIG11-GA-2012-322156; Spanish Ministry of Health (Instituto de Salud Carlos III) (grant PI14/00372); Bayer AG, From Targets to Drugs (grant 2015-03-1282) and Fundación FIPSE (grant 12-00001344-15). Ministerio de Economía y Competitividad, MINECO (grant CTQ-2015-68380-R) to J.C.M and grant SAF2015-63935R to M.G.L.; GW thanks Cancer Research UK (grant C9344/A10268), the BBSRC (grant BB/L01923X/1), the Bloomsbury Consortium and UCL School of Pharmacy for financial support. G.T. thanks Universidad Complutense for a predoctoral fellowship. P.M. thanks MECD for FPU fellowship (FPU13/03737). I. G. thanks UAM for a FPI fellowship. We also gratefully acknowledge the continued support of Instituto-Fundación Teófilo Hernando, Madrid, Spain.Peer Reviewe

High-Resolution Detection of Rock-Forming Minerals by Permittivity Measurements with a Near-Field Scanning Microwave Microscope

The identification of the minerals composing rocks and their dielectric characterization is essential for the utilization of microwave energy in the rock industry. This paper describes the use of a near-field scanning microwave microscope with enhanced sensitivity for non-invasive measurements of permittivity maps of rock specimens at the micrometer scale in non-contact mode. The microwave system comprises a near-field probe, an in-house single-port vectorial reflectometer, and all circuitry and software needed to make a stand-alone, portable instrument. The relationship between the resonance parameters of the near-field probe and the dielectric properties of materials was determined by a combination of classical cavity perturbation theory and an image charge model. The accuracy of this approach was validated by a comparison study with reference materials. The device was employed to determine the permittivity maps of a couple of igneous rock specimens with low-loss and high-loss minerals. The dielectric results were correlated with the minerals comprising the samples and compared with the dielectric results reported in the literature, with excellent agreements